Upper 2 panels: Colocalization of gene expression sites with nuclear pores. (A) Combination of anti-PfNup116 immuno-fluorescence staining with RNA-FISH using a probe against the active var gene expressed in 3D7-G7 clone (PF3D7_0412700). Nucleus stained by DAPI. Bar, 1 μm.(B) As in (A) with FISH probe against 18S ribosomal RNA. (C) Fraction of cells with an overlap of FISH and nuclear pore foci. Lower panel: Colocalization of PfSet10-HA with nuclear pores. Combination of anti-PfNup116 and anti-HA immunofluorescence staining on early trophozoite stage parasite expressing recombinant PfSet10-HA. Nucleus stained by DAPI. Bar, 1 μm.Guizetti J, Martins RM, Guadagnini S, Claes A, Scherf A. Nuclear pores and perinuclear expression sites of var and rDNA genes correspond to physically distinct regions in Plasmodium falciparum. Eukaryot Cell. 2013 12(5):697-702

Schematic representation of the Plasmodium falciparum nuclear architecture. A schematic representation of an infected red blood cell (RBC) and an enlargement of the P. falciparum nucleus are shown. From the centre towards the periphery we have defined three distinct areas (1–3) in which nuclear proteins specifically distribute. A summary is shown in which trophozoite stages have been analyzed. Proteins, previously predicted to functionally interact, are shown in red. Proteins previously predicted for their histone target sites are indicated by grey boxes. A, apicoplast. M, mitochondria, No, nucleolus.Volz J, Carvalho TG, Ralph SA, Gilson P, Thompson J, Tonkin CJ, Langer C, Crabb BS, Cowman AF. Potential epigenetic regulatory proteins localize to distinct nuclear sub-compartments in Plasmodium falciparum. Int J Parasitol. 2010 40(1):109-21. Copyright Elsevier.

Colocalization of histone H3 marks with PfSET10. Shown are from top to bottom: H3K4m1, H3K4m2, H3K4m3, and H3K9ac. The histone marks (red) were detected using specific antibodies and colocalized with PfSET10-HA (green) using anti-HA antibodies. Nucleus position is shown by DAPI staining. PfSET10 colocalized with the H3K4me1, me2, and me3 marks in the perinuclear region, which is also marked by H3K9ac Volz JC, Bártfai R, Petter M, Langer C, Josling GA, Tsuboi T, Schwach F, Baum J, Rayner JC, Stunnenberg HG, Duffy MF, Cowman AF. PfSET10, a Plasmodium falciparum methyltransferase, maintains the active var gene in a poised state during parasite division. Cell Host Microbe. 2012 11(1):7-18.

A. IFA-FISH of 3D7SET10-HA, in which var2csa (PFL0030c) activation has been selected by CSA adherence (‘‘on’’) (bottom panel). Unselected 3D7SET10-HA (top panel) has var2csa ‘‘off.’’ The localization of var2csa (green) and PfSET10 (red) is shown with both merged with the nucleus stained by DAPI (blue). Scale bars are equivalent to 0.5 mm. B. PfHP1 does not localize with active var2csa (‘‘on’’). The var2csa gene (red) location is shown with respect to PfHP1 protein (green). C. PfHP1 (red) and D. histone mark H3K9me3 (red,), known markers of silent var gene clusters and PfSET10 (green), do not colocalize in the nucleus stained with DAPI (blue).Volz JC, Bártfai R, Petter M, Langer C, Josling GA, Tsuboi T, Schwach F, Baum J, Rayner JC, Stunnenberg HG, Duffy MF, Cowman AF. PfSET10, a Plasmodium falciparum Methyltransferase, Maintains the Active var Gene in a Poised State during Parasite Division. Cell Host Microbe. 2012 11(1):7-18.

Identification of proteins that localize to the active var gene expression site. The table below shows proteins uniquely identified in the 3D7SET10-HA pull-down along with the number of unique peptides obtained. The PfIDs of the genes identified are shown along with their functional annotation.IFA of PfSET10 (green) and PfActin-1 (red) with the nucleus (blue) stained with DAPI. La R=late ring, Ea T=early trophozoite.Volz JC, Bártfai R, Petter M, Langer C, Josling GA, Tsuboi T, Schwach F, Baum J, Rayner JC, Stunnenberg HG, Duffy MF, Cowman AF. PfSET10, a Plasmodium falciparum Methyltransferase, Maintains the Active var Gene in a Poised State during Parasite Division. Cell Host Microbe. 2012 11(1):7-18.